Quantitative multianalyte microarray immunoassay utilizing upconverting phosphor technology

H. Päkkilä (Corresponding Author), M. Ylihärsilä (Corresponding Author), S. Lahtinen, Liisa Hattara, N. Salminen, R. Arppe, M. Lastusaari, Petri Saviranta, T. Soukka

Research output: Contribution to journalArticleScientificpeer-review

41 Citations (Scopus)

Abstract

A quantitative multianalyte immunoassay utilizing luminescent upconverting single-crystal nanoparticles as reporters on an antibody array-in-well platform was demonstrated. Upconverting nanoparticles are inorganic rare earth doped materials that have the unique feature of converting low energy infrared radiation into higher energy visible light. Autofluorescence, commonly limiting the sensitivity of fluorescence-based assays, can be completely eliminated with photon upconversion technology because the phenomenon does not occur in biological materials. Biotinylated antibodies for three analytes (prostate specific antigen, thyroid stimulating hormone, and luteinizing hormone) were printed in an array format onto the bottom of streptavidin-coated microtiter wells. Analyte dilutions were added to the wells, and the analytes were detected with antibody-coated upconverting nanoparticles. Binding of the upconverting nanoparticles was imaged with an anti-Stokes photoluminescence microwell imager, and the standard curves for each analyte were quantified from the selected spot areas of the images. Single analyte and reference assays were also carried out to compare with the results of the multianalyte assay. Multiplexing did not have an effect on the assay performance. This study demonstrates the feasibility of upconverting single-crystal nanoparticles for imaging-based detection of quantitative multianalyte assays.
Original languageEnglish
Pages (from-to)8628-8634
Number of pages6
JournalAnalytical Chemistry
Volume84
Issue number20
DOIs
Publication statusPublished - 2012
MoE publication typeA1 Journal article-refereed

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Microarrays
Phosphors
Assays
Nanoparticles
Antibodies
Single crystals
Streptavidin
Thyrotropin
Prostate-Specific Antigen
Luteinizing Hormone
Multiplexing
Image sensors
Biological materials
Rare earths
Dilution
Photoluminescence
Photons
Fluorescence
Infrared radiation
Imaging techniques

Cite this

Päkkilä, H., Ylihärsilä, M., Lahtinen, S., Hattara, L., Salminen, N., Arppe, R., ... Soukka, T. (2012). Quantitative multianalyte microarray immunoassay utilizing upconverting phosphor technology. Analytical Chemistry, 84(20), 8628-8634. https://doi.org/10.1021/ac301719p
Päkkilä, H. ; Ylihärsilä, M. ; Lahtinen, S. ; Hattara, Liisa ; Salminen, N. ; Arppe, R. ; Lastusaari, M. ; Saviranta, Petri ; Soukka, T. / Quantitative multianalyte microarray immunoassay utilizing upconverting phosphor technology. In: Analytical Chemistry. 2012 ; Vol. 84, No. 20. pp. 8628-8634.
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Päkkilä, H, Ylihärsilä, M, Lahtinen, S, Hattara, L, Salminen, N, Arppe, R, Lastusaari, M, Saviranta, P & Soukka, T 2012, 'Quantitative multianalyte microarray immunoassay utilizing upconverting phosphor technology', Analytical Chemistry, vol. 84, no. 20, pp. 8628-8634. https://doi.org/10.1021/ac301719p

Quantitative multianalyte microarray immunoassay utilizing upconverting phosphor technology. / Päkkilä, H. (Corresponding Author); Ylihärsilä, M. (Corresponding Author); Lahtinen, S.; Hattara, Liisa; Salminen, N.; Arppe, R.; Lastusaari, M.; Saviranta, Petri; Soukka, T.

In: Analytical Chemistry, Vol. 84, No. 20, 2012, p. 8628-8634.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Päkkilä, H.

AU - Ylihärsilä, M.

AU - Lahtinen, S.

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AU - Arppe, R.

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AU - Saviranta, Petri

AU - Soukka, T.

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AB - A quantitative multianalyte immunoassay utilizing luminescent upconverting single-crystal nanoparticles as reporters on an antibody array-in-well platform was demonstrated. Upconverting nanoparticles are inorganic rare earth doped materials that have the unique feature of converting low energy infrared radiation into higher energy visible light. Autofluorescence, commonly limiting the sensitivity of fluorescence-based assays, can be completely eliminated with photon upconversion technology because the phenomenon does not occur in biological materials. Biotinylated antibodies for three analytes (prostate specific antigen, thyroid stimulating hormone, and luteinizing hormone) were printed in an array format onto the bottom of streptavidin-coated microtiter wells. Analyte dilutions were added to the wells, and the analytes were detected with antibody-coated upconverting nanoparticles. Binding of the upconverting nanoparticles was imaged with an anti-Stokes photoluminescence microwell imager, and the standard curves for each analyte were quantified from the selected spot areas of the images. Single analyte and reference assays were also carried out to compare with the results of the multianalyte assay. Multiplexing did not have an effect on the assay performance. This study demonstrates the feasibility of upconverting single-crystal nanoparticles for imaging-based detection of quantitative multianalyte assays.

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JF - Analytical Chemistry

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Päkkilä H, Ylihärsilä M, Lahtinen S, Hattara L, Salminen N, Arppe R et al. Quantitative multianalyte microarray immunoassay utilizing upconverting phosphor technology. Analytical Chemistry. 2012;84(20):8628-8634. https://doi.org/10.1021/ac301719p